PrP expression in B lymphocytes is not required for prion neuroinvasion.

Prion diseases are typically initiated by infection of peripheral sites, as in the case of bovine spongiform encephalopathy, new variant Creutzfeldt-Jakob disease, kuru and most cases of iatrogenic Creutzfeldt-Jakob disease. In mouse scrapie, prion infectivity accumulates in lymphoid organs, and the absence of mature B lymphocytes prevents peripherally administered prions from inducing central nervous system disease. We have now assessed whether expression of the cellular prion protein, PrPc, is required for B lymphocytes to mediate neuroinvasion. We found that repopulation of SCID and Rag-1(-/-) mice with fetal liver cells from either PrP-expressing or PrP-deficient mice and from T-cell deficient mice, but not from B-cell deficient mice, is equally efficient in restoring neuroinvasion after intraperitoneal inoculation of scrapie prions. These results indicate that cells whose maturation depends on B cells or their products, such as follicular dendritic cells, may enhance neuroinvasion. Alternatively, B cells may transport prions to the nervous system by a PrP-independent mechanism.

Impaired prion replication in spleens of mice lacking functional follicular dendritic cells.

In scrapie-infected mice, prions are found associated with splenic but not circulating B and T lymphocytes and in the stroma, which contains follicular dendritic cells (FDCs). Formation and maintenance of mature FDCs require the presence of B cells expressing membrane-bound lymphotoxin-alpha/beta. Treatment of mice with soluble lymphotoxin-beta receptor results in the disappearance of mature FDCs from the spleen. We show that this treatment abolishes splenic prion accumulation and retards neuroinvasion after intraperitoneal scrapie inoculation. These data provide evidence that FDCs are the principal sites for prion replication in the spleen.

Use of brain grafts to study the pathogenesis of prion diseases.

For the study of prion neurotoxicity, we used neural-grafting techniques: mice devoid of the normal host prion protein (Prnp% mice) received a neural graft and were intracerebrally infected with mouse prions. The growth and differentiation properties of neural grafts were defined. Growth of embryonic neuroectodermal tissue was optimal at gestational days 12.5-13.5. The blood-brain barrier is reconstituted after 7 weeks in most animals. Scrapie-infected PrPC-expressing grafts develop a severe spongiform encephalopathy and contain proteinase-resistant protein and infectivity. Infected grafts deliver high amounts of prions to the host brain without eliciting disease. Infected grafts show a progressive disruption of the blood-brain barrier. Following intraocular prion inoculation of a transplanted Prnp% mouse, prions do not reach the intracerebral graft, indicating that PrP expression is required for propagation along the optic tract.

[Survival factors in the treatment of hereditary retinal degeneration]. / Uberlebensfaktoren in der Therapie erblicher Netzhautdegenerationen.

Hereditary retinal degeneration is characterized by apoptotic photoreceptor loss, a process governed by intricate molecular interplay and initiated when proapoptotic signals predominate in the individual cell. Identification of molecules involved and their actions has paved the way for testing the ones with anti-apoptotic functions in models of inherited retinal degeneration. Many of these factors are able to slow the course of the degeneration. However, to date no such treatment has been able to stop or even prevent the devolution of the disorder. Moreover, preservation of morphology does not necessarily correlate with preservation of ERG function. Deepened understanding of the pro- and anti-apoptotic networks is clearly needed for survival factors to be feasible for therapy in humans. In comparison, in a dog model of Leber's congenital amaurosis gene therapy could establish retinal function, thus supplying proof of efficacy of the method.

Transgenic and knockout mice in research on prion diseases.

Since the discovery of the prion protein (PrP) gene more than a decade ago, transgenetic investigations on the PrP gene have shaped the field of prion biology in an unprecedented way. Many questions regarding the role of PrP in susceptibility of an organism exposed to prions have been elucidated. For example mice with a targeted disruption of the PrP gene have allowed the demonstration that an organism that lacks PrPc is resistant to infection by prions. Reconstitution of these mice with mutant PrP genes allowed investigations on the structure-activity relationship of the PrP gene with regard to scrapie susceptibility. Unexpectedly, transgenic mice expressing PrP with specific amino-proximal truncations spontaneously develop a neurologic syndrome presenting with ataxia and cerebellar lesions. A distinct spontaneous neurologic phenotype was observed in mice with internal deletions in PrP. Using ectopic expression of PrP in PrP knockout mice has turned out to be a valuable approach towards the identification of host cells that are capable of replicating prions. Transgenic mice have also contributed to our understanding of the molecular basis of the species barrier for prions. Finally, the availability of PrP knockout mice and transgenic mice overexpressing PrP allows selective reconstitution experiments aimed at expressing PrP in neurografts or in specific populations of hemato- and lymphopoietic cells. Such studies have shed new light onto the mechanisms of prion spread and disease pathogenesis.

Intramedullary pressure increase for different commercial and experimental reaming systems: an experimental investigation.

OBJECTIVE: To measure the differences in intramedullary (IM) pressure for commercial reamer systems. DESIGN: IM pressure values for the following systems were measured: AO, Biomet, Howmedica grey reamer, Richards, and Zimmer. To investigate the influence of shaft diameter, the AO reamer head was additionally connected to a small shaft (A6/A7). The pressures were measured in plexiglass tubes filled with a mixture of petroleum jelly and paraffin oil with flow properties at 20 degrees C equivalent to those of bovine medullary fat at 36 degrees C. The reaming assemblies were inserted into the tubes using a materials testing machine at a constant speed. In addition, pressure measurements were made using five pairs of human femora to compare Biomet reamers with the AO reamer with thin, flexible drives (A6/A7). RESULTS: The following pressure distributions were obtained (millimeters of mercury; mean value +/- standard deviation): 9.5-millimeter reamer: low for Biomet (272+/-39); moderate for Richards (810+/-101); and high for Howmedica (990+/-132), AO conventional (1,000+/-97), and Zimmer (1,140+/-183); 13.0-millimeter reamer: low for Biomet (132+/-21), Howmedica (204+/-45), and Zimmer (226+/-33); moderate for AO conventional (474+/-42); and very high for Richards (1,734+/-127). The second worst system (AO conventional: 1,000+/-97) became the second best system by simple reduction of the shaft diameter (A6/A7: 378+/-33). CONCLUSION: A comparison of shaft diameters and pressure increase clearly showed that the system with the thinnest shaft produced the lowest pressure values and vice versa.

[The AO external clamp fixator. New techniques in external stabilization of tibial fractures]. / Zangenfixateur externe der AO. Neue Techniken bei der externen Stabilisierung von Unterschenkelfrakturen.

A serious disadvantage of conventional external fixators is the opening of the medullary canal. This creates a direct communication between the canal and the exterior. The new pinless external fixator does not have this disadvantage. The clamps of the new fixator system do not penetrate cortical bone, they simply rest on the cortical surface. The clinical indications of this easily managed system are fractures of the tibial shaft in cases in which primary internal fixation is precluded either by precarious local or general conditions, or even by a lack of infrastructure. In such situations the pinless external fixator is an excellent device to achieve a good fracture stabilisation rapidly, while leaving open all subsequent alteration of treatment. In case of secondary change to an intramedullary nail the pinless fixator facilitates the procedure, because this fixator can be also used as an distractor during nailing. Furthermore this fixator can be combined with any intramedullary nail to increase the stability in case of very proximal or distal as well as highly instable shaft fractures. The clamps of the pinless external fixator set are used in combination with the standard AO fixator system.

[Innovative implants -- a prerequisite for biological osteosynthesis]. / Neue Implantate als Voraussetzung biologischer Osteosynthesen.

Biological osteosynthesis is a new paradigm in fracture fixation. In contrast to the conservative mechanical operative period, where exact anatomical and rigid fracture fixation was the ultimate ambition, the new perception in fracture management attaches more importance to the conservation of bone perfusion than to the exact anatomical reduction. Because soft tissue condition and local blood circulation are most important for bone reconstruction and healing of fractures, additional damage of soft tissue caused by operation should ideally be avoided. Consequently surgeons aim at minimally invasive operating techniques using no-touch bone implants. This shift in point of view has also been accommodated by a rapid progress in development of innovative bone implants. Solid medullar nails, novel designs and applications of plates, locking screws and biocompatible titanium implants are some of the implantation techniques which pushed the new concept of biological fracture fixation.

[Locking of the intramedullary nail in the medullary cavity of the tibia (author's transl)]. / Die Verklemmung des Marknagels in der Markhöhle der Tibia. In vitro-Messung der Längs- und Querdeformation des Marknagels mit Hilfe von Dehnungs-messstreifen.

Whether or not an effective locking of the medullary nail in transverse direction is achieved after medullary nailing of the tibia, is a moot point in literature. Researchers agree, however, that the essential locking of the nail occurs on the longitudinal axis of the nail. The authors conducted studies with strain gauges measuring the expansion and introduced together with the nail in order to obtain information on the elastic performance of the nail in transverse and longitudinal direction. The curves plotted from the data obtained, present a typical appearance. After the tibia has been properly reamed, a medullary nail of the AO is subjected to both elastic transverse compression as well as slight longitudinal bending after nailing. In case of excessive reaming the transverse elastic effect is only slight. If, on the other hand, reaming is not effected wide enough, the bone may burst open or the nail may undergo plastic deformation.

Locking Compression Plate (LCP). An osteosynthesis plate based on the Dynamic Compression Plate and the Point Contact Fixator (PC-Fix).

Based on many years of experience with compression plating and promising results obtained with so-called internal fixators, an implant system was developed which combines the two treatment methods in one implant. Despite the combination of these different fracture treatment techniques, no compromises were made with regard to application as a compression plate or as a bridging device in the form of an internal fixator. The individual construction elements in this new implant system have individually proven themselves extremely valuable in clinical practice. The integration of these well proven elements into the new implant system has made it compatible with the majority of existing instruments and conventional screws. The surgeon is free to select the best treatment method to suit the fracture situation and to make combinations as and when necessary. The LCP (Locking Compression Plate), the product of these combinations, is in line with the latest plating techniques, the aim of which is to achieve the smallest possible surgical incisions and to preserve the blood supply to the bone and the adjacent soft tissues.

Effect of flexible drive diameter and reamer design on the increase of pressure in the medullary cavity during reaming.

Reaming the medullary cavity before insertion of an intramedullary nail, results in an increase in both temperature and pressure. This may lead to aseptic necrosis of the cortex and to fat embolism, whereby the increased pressure in the medullary cavity plays a significant role. This paper aims to determine whether a reduction of the drive diameter combined with a different reamer design reduces the amount of intramedullary pressure. The pressure levels were measured in plexiglass tubes filled with a mixture of vaseline and paraffin. The viscoelastic properties of this mixture at 20 degrees C were equivalent to those of bovine medullary fat at 36 degrees C. In comparison with the conventional reamer system (9.0 mm drive + 9.5 mm AO reamer), the 7.0 mm drive+AO reamer and the 7.0 mm drive+hollow reamer, the pressure values were reduced as follows: 1. 9.0 mm drive + 9.5 mm hollow reamer: diaphyseally by 19%, metaphyseally by 21% 2. 7.0 mm drive + 9.5 mm AO reamer: diaphyseally by 48%, metaphyseally by 49% 3. 7.0 mm drive + 9.5 mm hollow reamer: diaphyseally by 61%, metaphyseally by 66%. If the gap between the flexible shaft and the wall of the plexiglass tube became large enough, only small pressure values were recorded for all three types of reamer. In summary, it can be stated that the reduction of the drive diameter causes a sustained reduction in the intramedullary pressure, but that the newly developed hollow reamer only leads to a further reduction in pressure in combination with a thin flexible drive.

[External fixator: surgical technique, pinless fixator, change in procedure]. / Fixateur externe: Operationstechnik, pinless fixator, Verfahrenswechsel.

External Fixation-Technique: The advantages of external over internal fixation are as follows: a) endosteal and periosteal blood supply is undisturbed, b) "low-tech" equipment may be used, c) secondary adjustments are possible and d) easy implant removal. These benefits however are outweighed by the main disadvantages of long term external fixation i.e. pin complications and delayed union of fractures. Better understanding of postoperative management and careful application of screws of improved design will lead to better results. Today's standard applications of external fixation for tibial fractures is a unilateral fixator, using Schanz screws. The pin-bone interface is the most critical site of all external fixation. By avoiding heat necrosis (low temperature drilling) and preventing micro motion at the pin-bone interface (by applying bending- or more recently radial-preload), pin complications such as infection and loosening can be reduced. Two Schanz screws are inserted into each main fragment and are connected with one short tube per fragment. The fracture is then reduced by using these tubes as handles. After reduction a third tube connects the first two by means of two tube-to-tube clamps. This type of fixation will easily allow for three dimensional secondary corrections of alignment. Approximately three weeks following the injury some motion at the fracture site will stimulate callus formation. This can be achieved by destabilisation, dynamisation or "active stimulation" of the fracture site [2]. Pinless fixator: The pinless external fixator holds the fragments firmly with pointed clamps that penetrate about one millimeter into cortical bone without entering and contaminating the medullary canal.(ABSTRACT TRUNCATED AT 250 WORDS)

Depth-force patterns of periodontal probing. Attachment-gain in relation to probing force.

It was the purpose of this study to determine whether probing force had an influence on the amount of clinical attachment-gain assessed after treatment by scaling and rootplaning. A probing device was constructed which allowed simultaneous monitoring of probing force and probe penetration and which standardized the insertion pathway for repeated measurements. In 10 periodontal patients, 2 deep pockets were selected which were measured before and after periodontal treatment by scaling and root-planing. Depth-force plots were compared by superimposition. Depth values were determined at 5 different force levels (0.25, 0.50, 0.75, 1.00 and 1.25 N) on each plot and changes of clinical attachment levels were calculated. A significant relationship was seen between probing force and attachment level. The values obtained with 0.25 N were significantly different from the values obtained with higher forces (p less than 0.001). Slight, but non-significant differences were noted in the amount of attachment-gain obtained at the 5 force levels. At a probing force level of 0.25 N, there was 0.80 mm mean attachment gain. With 0.50 N, there was a gain of 0.70 mm; with 0.75 N the gain amounted to 0.67 mm in mean. At 1.00 N and at 1.25 N, a gain of 0.66 mm was recorded.

[Epiperiostal, percutaneous plate osteosynthesis. A new minimally invasive technique with reference to "biological osteosynthesis"]. / Die epiperiostale, perkutane Plattenosteosynthese. Eine minimal-invasive Technik unter dem Aspekt der "biologischen Osteosynthese".

Conservation of bone perfusion, protection of the soft tissue envelope and reduction of systemic stress by strengthening the host defence mechanism are general and essential aspects of a biological osteosynthesis. The minimal invasive operating techniques with the use of technical aids and tricks form the necessary presupposition for successful bone healing with a low complication rate. For an epiperiosteal, percutaneous plate osteosynthesis, the technique using a sliding tip and a manipulation handle is demonstrated.

Biomechanics of the interlocking nail. A study of the proximal interlock.

Conventional interlocking nails for the femur use an inclined proximal interlocking screw; the AO/ASIF universal interlocking nail utilizes a transverse interlocking screw which is anchored in cortical bone. The transverse position of the screw allows one to employ the nail for either the right or left side. The yield strength of the thinner diameter transverse screw, (thread 4.5 mm. core 3.0 mm) was compared with that of the Grosse-Kempf nail oblique screw (thread 6.3 mm, core 4.5 mm) by testing in cadaver femora to evaluate the risk of fatigue failure. The test, performed using slow loading rates (50 N/s), simulated either a diaphyseal or a subtrochanteric fracture situation. The test rig applied a load corresponding to body weight and pull at the trochanter. Taking the highest load without plastic deformation as a criterion, the mechanical strength of the two screws was evaluated for each fracture situation. The thinner but shorter transverse interlocking screw demonstrated higher strength (4500 N vs. 3000 N; P less than or equal to 0.02) than the thicker inclined screw in the diaphyseal and the subtrochanteric fracture situation (2300 N vs. 1100 N; P less than or equal to 0.06). The biomechanical test showed the transverse screw to have sufficient and superior strength.

Biomechanical evaluation of the Pinless external fixator.

In open fractures especially in those with severe soft tissue damage, fracture stabilisation is best achieved by using external fixators. There are some intrinsic complications which occur during classical external pin fixation. To overcome the problem of pin track infection and vascular damage from drilling, the Pinless external fixator was developed. It is based on the idea of a forceps with trocar points, which only penetrate the bone cortex superficially. The function of the device was tested in two mechanical trials and two in vitro tests in which one pinless clamp was put under a controlled load of 50 N, 150 cycles/day and studied over a 5 week period in sheep. The loads and time range of the experiment were chosen to simulate a temporary fracture stabilisation in a patient not bearing weight. The main question to be answered was whether the Pinless external fixator would be able to maintain stable fixation. Furthermore, it was to determine the changes at the trocar-to-bone interface. The clamp was found to maintain 72% of the initially applied clamping force after 5 weeks of in vivo application and it was found to be tight at removal. Some decrease of clamping force was found during the first 20 days and then the force tended to level off. There was no slippage nor did the clamp penetrate the cortex. There were no obvious signs of infection around the trocar-holes and in the bacterial tests no pathological cultures were grown. Histology revealed very localised bone reactions, the indentation caused by the trocar tips being only 1.2 mm deep. The study concludes, as far as could be ascertained from these tests, that it is safe to use pinless external fixation for temporary fracture fixation.

[Unreamed tibial nail in tibial shaft fractures with severe soft tissue damage. Initial clinical experiences]. / Der unaufgebohrte Tibianagel (UTN) bei Unterschenkelschaftfrakturen mit schwerem Weichteilschaden. Erste klinische Erfahrungen.

In a prospective study, since March 1989, 55 tibial shaft fractures have been treated with a new, unreamed solid tibial nail (UTN). This nail was initially designed as a temporary implant. The first 33 cases with second or third degree soft tissue damage were reviewed 6 months or more after the operation. Fractures were classified according to Müller: 6 type A (18.2%), 15 type B (45.5%), and 12 type C (36.7%). In 9 cases (27.3%), there was GII (n = 4) or GIII (n = 5) closed soft tissue damage according to Tscherne's classification. The 24 open fractures (72.7%) comprised 11 OII, 3 OIIIA and 10 OIIIB fractures (Gustilo classification). 24 patients (72.7%) were polytraumatized, the mean PTS (Hannover Polytrauma Score) was 18 points (range: 8-65 points). All fractures were stabilized without reaming. The implant diameter was 8mm (n = 14) or 9 mm (n = 19). Static locking was performed in 31 cases. Dermatofasciotomy was necessary because of compartment syndrome in 14 cases. In 1 grade IIIB open fracture soft tissue coverage was performed with a latissimus dorsi myocutaneous free flap 4 days after nailing. In 32 of the 33 cases the use of an additional cast or brace was not necessary during the follow-up treatment; 1 patient had a cast for 8 weeks for the treatment of accompanying injuries. Full weight-bearing was achieved in 5 cases within 3 weeks, in 16 cases within 12 weeks, and in 30 cases within 26 weeks. In 16 cases (48.5%) the interlocking screws were removed after 5-26 weeks (mean: 10 weeks).(ABSTRACT TRUNCATED AT 250 WORDS)

The development of the distal femur Less Invasive Stabilization System (LISS).

Fractures around the knee typically require operative fixation to achieve an acceptable, functional outcome. The idea behind the Less Invasive Stabilization System (LISS) was to combine the advantages of both interlocked intramedullary nailing techniques and the early advances of the so-called biological plating technique into one system. This paper introduces the mechanical concept of a locked internal fixator and details some important aspects of the anatomical and biomechanical development of the LISS.

Reaming versus non-reaming in medullary nailing: interference with cortical circulation of the canine tibia.

Medullary nails have only been used with reluctance in the treatment of open fractures since reaming interferes with the cortical circulation. The technique of interlocking offers the interesting possibility of inserting a relatively stiff nail which does not necessarily require reaming. The effects of reaming prior to nail insertion upon the cortical circulation was investigated in the canine tibia. Intravital staining with procion red was used to trace areas of disturbed circulation. Nail insertion without reaming provided clear advantages for the bone's blood supply, while reaming disturbed perfusion in two-thirds of the cortical area and regionally extended through the entire thickness of the cortex. The disturbance without reaming was limited to the inner layer of the cortex and involved only one-third of the cortical cross-section.

The hybrid ring tubular external fixator: a biomechanical study.

OBJECTIVE: To measure and compare the mechanical properties in bending of the four-ring, and three-ring/one-tube hybrid external fixation frames. DESIGN. IN VITRO: measurements of the mechanical behaviour of ring and ring-tubular external fixation frames. In the latter, one ring of the full circular frame was replaced by one tube and Schanz screws. BACKGROUND: The mechanical properties of the classical Ilizarov four-ring external fixation frames has been compared to those of other external fixation frames by various authors. However, in clinical practice the hybrid fixation frame is being used with increasing frequency. Therefore the mechanical properties of the latter are of immediate interest and clinical value. METHODS: On explanted sheep tibiae with single and double osteotomies, frame stiffness in the four-point bending mode was measured at different K-wire tensions, comparing the values obtained from four-ring frames, to those of three-ring-tubular hybrid frames. These measurements were made under conditions of (a) bone distraction (BD), and (b) segment transport (ST), both at the initial and final stages of this procedure. RESULTS: In circular frames, frame stiffness in bending for increasing K-wire tension showed a Gaussian distribution both in distraction and post-ST with an optimum at 1000 N. In ring tubular hybrid frames, however, frame stiffness showed a more linear relationship to K-wire tension. CONCLUSIONS: In the four-ring Ilizarov external fixation frame, the exchange of one ring with one tube and one Schanz screw both reduced frame stiffness in bending and converted to linear its relationship to K-wire tension. RELEVANCE: Under clinical conditions, the use of a similar ring tubular hybrid external fixator allows the adjustment of frame stiffness in a simple and practical way. This is not the case with the original ring fixation frame.